First strand cDNA for the PCR amplification was prepared from 0.5 µg total RNA using PrimeScript™ RT Reagent Kit - Perfect Real Time (Takara). cDNA was diluted 1:40 and 5 μl were used for each quantitative polymerase chain reaction (qPCR) performed on 96-well plates, using ...
A t-test was used between two groups. The two-way ANOVA was conducted for the factorial design experiments. A P-value less than 0.05 was considered statistically significant. The criteria using the F and P values to determine the interaction type were that additive effect met F < 5, P >...
Subsequently, cell lysates were clarified by centrifugation (15,000 g, 20 min, 4 °C) and protein concentration was determined using the Pierce™ BCA Protein Assay Kit (Thermo Scientific). Aliquots containing 20 μg of total cellular proteins were denatured in Laemmli buffer (50 mM DTT, ...